Artificial spawning and larvae performance of

three Indonesian mahseer species

1,2
Wahyulia Cahyanti, 2Dinar T. Soelistyowati, 2Odang Carman,
1
Anang H. Kristanto
1
Research Institute for Freshwater Aquaculture and Fisheries Extension, Jl. Sempur
No. 1, Bogor, Indonesia; 2 Aquaculture Department, Bogor Agricultural University,
Dramaga Campus, Bogor 16680, West Java, Indonesia. Corresponding author:
W. Cahyanti, wahyulia_kkp@yahoo.com

Abstract. This study attempts to evaluate the reproductive and larvae performance of the three species
of mahseer in Indonesia through artificial propagation. The research was conducted at the Research
Institute for Freshwater Aquaculture and Fisheries Extension, Bogor, Indonesia from August to December
2017. A completely randomized design was performed using three mahseer species (Tor douronensis,
Tor soro and Tor tambroides) with three replications. The fecundity and egg diameter of Tor douronensis
were (2.3±0.07 mm; 4987.80±17.25 eggs kg-1), Tor soro (2.86±0.08 mm; 3157.02±97.28 eggs kg-1)
and Tor tambroides (3.00±0.06 mm; 1061.19±35.18 eggs kg-1) respectively. The fertilization rate (FR)
and hatching rate (HR) between species ranged between 76.00±7.21 and 93.33±1.15 (FR) and
80.36±6.00% to 93.32±4.57 (HR), while the survival rate of 15-day-old larvae was not significantly
different between Tor soro and Tor tambroides, but significantly different with Tor douronensis.
Key Words: breeding, eggs, larvae, mahseer, reproduction.

Introduction. Four species of mahseer (genus Tor) were identified to be endangered,

i.e., Tor douronensis, Tor soro, Tor tambra and Tor tambroides (Robert 1999),
indigenous from Sumatra, Java and Kalimantan islands due to limited cultivation activity
knowledge (Kunlapapuk & Kulabtong 2011) and environmental degradation (Asih et al
2008). Recognizing the importance of socio economic fisheries and protection of these
natural stocks, the development of reproduction technology on aquaculture is necessary
for sustainable production (Abderrazik et al 2016). The basic information on the potential
of fish reproduction can be obtained from the gonadal maturation review until the fish
spawn and produce larvae (Setyaningrum & Wibowo 2016).
Mahseer has been successfully domesticated in Indonesia. The breeding
technology of Tor soro was declared successful in 2010 (Farastuti et al 2014). Indeed,
the reproductive and larvae performance of mahseer needs to be developed to improve
its production. Dunham (2004) states that fish from different locations have different
reproductive characteristics related to the influence of interactions between their genetics
and the environment. Generally, mahseer spawns naturally (Dinesh et al 2010, Redjeki
2007; Nautiyal 2014); however, Abderrazik et al (2016) conducted research on
reproductive biology knowledge to determine the genetic and reproductive potential of
Sardina pilchardus. A study on artificial spawning between existing mahseer species in
Indonesia has never been done. The nutritional requirement and the condition of the
environment both determine how broodstock is reared and maintained for gonad
maturation. In this framework, the present study attempts to obtain baseline data related
to reproductive performance and the larvae of three species of Tor produced by artificial
spawning.

Material and Method. This reasearch was conducted from November 2017 to January
2018. Three species of Tor genus from the Research Institute for Freshwater Aquaculture
and Fisheries Extension, (Bogor, Indonesia) were used as broodstock. The size of female

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broodstock ranged 40-62 cm total length (TL) and 940-4880 g in body weight (BW),
while the male broodstock ranged 30-45 cm (TL) and 500-990 g in (BW). Five pairs of
each species were acclimatized in ponds for approximately one month; they continued for
gonad maturation, spawning and larvae rearing. Gonad maturation for each species was
carried out by separately rearing between males and females in a 200 x 300 x 70 cm
pond, carrying an artificial feeding containing 28% of protein with a 3% feeding rate per
day (twice daily, morning and afternoon). Gonad maturation was measured every two
weeks through the cannulation method.
The fish broodstock with mature gonads was injected with LHRH-a + anti-
dopamine hormone at dosage 0.6 mL kg-1 of body weight (one injection at 13.00 pm and
one at 17.00 pm, then stripped after a day). A gram of eggs was sampled and the egg
number counted as the relative fecundity (eggs kg-1) based on gravimetric analysis.
Furthermore, the remaining eggs were fertilized with sperm and taken into the incubation
in three aquariums with sizing 30 x 40 x 20 cm (500 eggs/aquarium). Each aquarium
was equipped with the recirculating water system and a moderate aeration system. Each
aquarium was stocked with 250 larvae fish from each species and was replicated three
times. The larvae were reared without being fed until the yolk disappeared, but
eventually fed at-satiation with Artemia nauplii (morning, afternoon and evening). Water
quality management was done by water exchange (50%) using a siphoning method. The
parameters measured consisted of fertilization rate (%), hatching rate (%) and
abnormality (%) of the 15 days old larvae. The gonadal maturity was analyzed
descriptively and the other parameters (egg diameters, time hatched, fertilization rate
(FR), hatching rate (HR), abnormality and survival rate (SR)) were analyzed by ANOVA
(p < 0.05). The water quality parameters including temperature, pH, dissolved oxygen
(DO) and ammonia concentration were measured on day 0, 15, 30, 45 and 60 in-situ at 8
and 10 am.

Results. Data of the three broodstock mahseer, eggs and fish larvae is presented in
Table 1.
Table 1
Data of three mahseer broodstock, eggs and their larvae

Parameter Tor douronensis Tor soro Tor tambroides

Mature female (induce to ovulate)
Total length / TL (mm) 405-435 460-520 516-600
Body weight / BW (g) 820-1020 930-1360 3580-4880
Number of female (individual) 5 5 5
Spawned fish (individual) 2 2 2
Atresia egg fish (individual) - 1 2
Mature male
Total length (mm) 355-450 299-318 300-342
Body weight (g) 460-990 290-300 300-390
Number of male 5 5 5
Spawned 5 5 4
Breeding
Latency period (hour) ±18 ±18 ±16
Eggs
Diameters of stripped eggs (mm) 2.3±0.07a 2.86±0.08b 3.00±0.06c
Total weight of stripped eggs (g) 40-50 50-70 70-100
Time hatched (day) 4.34±0.03a 5.02±0.02b 5.97±0.02c
Larvae
Abnormality (%) 8.40±1.83a 2.47±0.12b 2.93±0.31b
Survival rate (%) 79.33±0.95a 82.11±1.43b 80.22±1.05b
Some data presented are averages of ±SD from three replicates. Different superscripts in the different columns
show significant differences (p< 0.05).

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Egg diameter. The average egg diameter among species at the time of spawning was
2.3±0.07 mm (T. douronensis), 2.86±0.08 mm (T. soro) and 3.00±0.06 mm (T.
tambroides) (Figure 1). The egg diameter of the T. douronensis was smaller than others,
while T. soro and T. tambroides were not different (Figure 2).

Figure 1. Average egg diameter of the three Indonesian mahseer species.

Figure 2. Distribution of egg diameters at the time spawning of the three Indonesian
mahseer species.

Fecundity. The number of eggs (fecundity) among species was significantly different (p
< 0.05), 4987.80±17.25 eggs kg-1 (T. douronensis), 3157.02±97.28 eggs kg-1 (T. soro)
and 1061.19±35.18 eggs kg-1 (T. tambroides) respectively. The data refers to the
weights and lengths of successful spawning broodstock.

Fertilization and hatching rates. The hatching and fertilization rates in the Tor
douronensis were significantly lower than the other two species (Table 2). Tor
douronensis had the shortest domestication period of the three. The values of the
fertilization and hatching rates, in this study, are presented in Table 2.

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 Table 2
Fertilization and Hatching rates of three mahseer species

Species Fertilization rate Hatching rate

Tor douronensis 76.00±7.21a 80.36±6.00a
Tor soro 93.33±1.15b 93.32±4.57b
Tor tambroides 91.33±1.15b 90.20±2.75b
The data presented are averages±SD from three replicates. Different superscripts in the same columns show
significant differences (p < 0.05).

Analysis of water quality parameters. The water quality data during egg incubation
and larval rearing is presented in Table 3.

Table 3
Water quality parameters of media for egg incubation and larval rearing

Values
Water quality parameters
Present study Other studies
Water color Clear Clear*
pH 7.91-8.12 6.81-7.09**
Temperature (°C) 23.58-23.68 25.26-27.30**
DO (mg L-1) 7.16-7.30 6.30-8.34**
Nitrate (mg L-1) - 1.46-3.10***
Nitrite (mg L-1) - 0.004-0.2***
Ammonia (mg L-1) 0.08 _

(*) Kunlapapuk & Kulabtong (2011); (**) Rachmatika et al (2005); (***) Haryono & Subagja (2008).

Discussion. Feeding serves as a source of energy that will be used to sustain life,
growth and the reproductive process. The time required for the process of gonad
maturation up to spawning varies for each species, depending on the type of feed. The
present study showed that mahseer fed with 28% protein of feed attained gonad
maturity during 4-6 weeks. In this study, the gonad ripe response of T. tambroides was
faster than the other two species. This was possibly due to the fact that T. tambroides
has a relatively smaller protein requirement than T. soro and T. douronensis. According
to Kamler (1992), protein is the dominant component in egg yolk, while the amount and
composition of the protein is determined by the egg size. Mahseer fish, in the present
study, showed natural changes in shape, through the differences in gonad maturation
(abdomen). In general, the fish increased voluptuous gain, especially the inside part.
However, the belly returned to its regular shape once the fish was ready for ovulation,
marked by an increment volume and circumference of the abdominal cavity. A research
conducted by Hardjamulia et al (1999) revealed that feeding of T. douronensis with 36%
protein of feed led to the fish reaching gonad maturity within three weeks (21 days).
According to Redjeki et al (1999) on the previous research, Tor soro fed with 35%
protein of feed could reach maturity within 3-4 weeks and the abdominal cavity length of
Kancra fish mature ranges 29.0–34.5 cm (Redjeki 2007).
Latency time of the Tor genus was between 16-18 hours after the second injection
with water temperature 24-25oC. The latency duration was defined as the time between
injection and stripping. During this period, fish ovulation was examined up to two times.
The same dose of hormone 0.6 mL kg-1 was used, which caused the different fecundity
and latency time. These results reinforce the assumption that the fewer eggs produced,
the faster the latency time. Kristanto et al (2007) said that the range of latency time of
mahseer fish (T. soro) from the last hormone injection was 12-19 hours, at water
temperature 24-28oC; this latency time is necessary to know, because if it passes, the
fish eggs will become atresia.
The broodstock were not all spawned in each species. The T. tambroides male and
female spawned about 80%. It is suspected that this happened because the fish was sick,
seen from its dark color and slow movement. The T. douronensis female was only 40%

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successfully spawned, while T. soro was 60%. It was suspected that the nutritional
requirement of females in each species was different, while the fish were given the same
feed protein. Feed is an essential component needed for reproduction. The protein need
was different for each species and generally ranges from 25 to 40%.
The egg diameter differences were allegedly caused by genetic (non-additive) and
genetic-environment interaction (GE) factors. Each broodstock comes from a different
environment (province). This is considered to affect the physical and reproductive fish.
Falconer & Mackay (1996) stated that genotype by environment (G × E) interaction
refers to the difference in the response of genotypes to different environments. In
nature, the egg diameters of T. douronensis ranged from 2.22 to 2.49 mm (Gaffar et al
1991), while the egg diameters of Labeobarbus douronensis were 3.5±0.01334 mm
(Redjeki et al 2001). The range of eggs’ diameter varies, possibly for differences in gonad
maturity and time required for the maturation process of each fish species. The
difference is as a result of the process of vitellogenesis, hydration and grains formation of
oil that runs gradually. The more uniform the egg diameter, the better the quality of
spawning (Sivakumaran et al 2003). Different vitellogenesis processes are strongly
suspected to be influenced by generalized feeding, whereas the nutrient needs of each
species are not yet known.
The fecundity value of this type of fish (mahseer) still varied and could not be
ascertained. For example, T. tambroides with an average broodstock body weight of
5200-8700 g produced 3125-8201 eggs at the time of spawning (Haryono 2006), while
T. douronensis of 85.5 cm body length produced 63,360 eggs (Gaffar et al 1991) and
those with a body length of 64.8 cm produced 14,433 eggs (Rupawan et al 1999). The
fecundity of T. soro with the average body weight of 800-1000 g was 1752-2129 eggs
(Farastuti et al 2014) and T. tambroides with a body weight of 2230-2850 grams had
produced 1200-2800 eggs (Azuadi et al 2013). Gaffar et al (1991) mention T.
douronensis fish in South Sumatra, which recorded 2073 eggs kg-1. According to
Bromage et al (1993), an important factor affecting eggs (number and size) is the size of
the parent used. For the same age fish as in this study (2-3 year old): the greater the
weight/size of the parent, the higher the value of fecundity. In total, the number of eggs
produced is also in tune with the genetic diversity of each parent. The fecundity varied
among the species and individual, and has been known to be dependent on brood
conditions such as size (length, weight and age), genetics, food availability and
environmental factors (Muchlisin et al 2011). The fecundity of large size fishes is linearly
related to the total length of the fish (Joshi et al 2018).
At air temperatures of 23.58-23.68oC T. douronensis hatched at the fastest
(4.34±0.03 days), followed by T. soro (5.02±0.02 days) and T. tambroides (5.97±0.02
days). In general, Tor spp. hatch after 4-5 days of incubation at 25-27°C (Asih & Subagja
2003). Subagja et al (2013) stated that at a temperature of 25-26oC, T. douronensis
hatchery requires 72 hours with a hatching rate of 65%. Azuadi et al (2013) stated that
the hatching rate of T. tambroides eggs was resolved between 57.8-82.5% at a
temperature of 25-27oC. The hatch time in this study is indeed slower, this is due to the
lower incubation temperature. Even though the time needed to hatch is longer, but the
HR and SR generated reach 90% more.
The fertilization rate in fish is largely determined by the quality of eggs,
spermatozoa, media and human handling. Fish eggs will usually develop normally if the
hatchery conditions include oxygen, temperature and pH are fulfilled. In addition
hormones also give effect (Woynarovich & Horvath 1980). The use of hormones (sGnRHa
+ dopamine) not only encourages the female parent to ovulate alone, but also its
decision with successful fertilization, hatching and larvae produced (I'tishom 2008).
The 15-days old survival rate (SR) showed no significant difference among species
(p > 0.05). The survival rates of larvae T. douronensis, T. soro and T. tambroides were
79.33±0.95%, 82.11±1.43% and 80.22±1.05% respectively. This result was relevant
with Subagja et al (2009); the maintenance of T. douronensis for 88 days had SR of
80%. The survival rate of T. tambroides ranged between 48.9 and 84.2% (Azuadi et al
2013). A high SR value can be ascertained if the number of larvae produced in mahseer
is spawning quite well. So this type of fish is very good as aquaculture fish.

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 The bigest abnormality (8.40±1.83%) was observed on T. douronensis. This value
was significantly different (p > 0.05) than that of the other two species. Many factors
cause abnormality in fish larvae, including genetic factors on Sparus aurata (Fernandez et
al 2008), nutrients deficiency such as phosphorus on Melanogrammus aeglefinus, vitamin
C on Oncorhynchus mykiss and Ictalurus punctatus, vitamin A on Sparus aurata, vitamin
K on Melanogrammus aeglefinus (Berillis 2015), heavy metal pollution on Nile tilapia
(Hassanain et al 2012), water temperature on Sparus aurata (Georgakopoulou et al
2010) and sea bass Dicentrarchus labrax (Sfakianakis et al 2006) and a possible role of
bacteria and parasites of many fishes in the Willamette River (Kent et al 2004).
In this study, the larvae abnormalities were suspected from genetic suits for eggs
and larvae. Larvae are maintained in a controlled environment. Observation of
abnormalities was done visually. Commonly acquired disorders were spinal defects such
as kyphosis (bent upward), lordosis (crooked down) and scoliosis (crooked up and down /
like S). Skeletal deformities are major factors that downgrade hatcheries production. The
abnormality value obtained was relatively small; it was in line with Ismi et al (2007)
where the percentage of abnormality was visually lower than that of the staining method,
due to fish that visually looked normal, when they, in fact were defective. There is no
much research on larval abnormalities in artificial spawning of mahseer. Yuliyanti et al
(2016) researched on T. tambroides with different hatching temperature variations,
showing a value between 2 and 10%.
The water quality has affected fertilization and hatching rate. In this study, the
temperature was relatively lower than that in the natural conditions, although, the
temperature was still suitable for the fish species. However, other water quality
parameters exist in a good range for egg hatching and larval rearing. Management
activities to maintain proper water quality are essential in egg hatching and larval
rearing; a simple way is to use a recirculation system with a filter so that water quality
remains stable.
Based on data, Tor douronensis has reproductive performance less good than the
other two species. This is allegedly due to gonad maturity level in Tor douronensis at the
lowest spawning time. The low level of gonad maturity in Tor douronensis is evidenced by
the amount of fish that successfully spawn (two out of five individuals) and has the
smallest egg diameters. Good level of gonadal maturity can be predicted from the
number of eggs with a diameter above the average. Generally, the T. soro is the species
with the best reproductive potential compared to the other two. This is evident from the
uniformity of ovulated eggs size, FR, HR and SR.
The highest fecundity was in T. douronensis, so it was likely to produce many
larvae, but had the lowest survival rate with the highest larval abnormality. The fish
species that had high fecundity, high survival rate and low abnormality was T. soro. The
egg diameter of the three species is related to fecundity; it was in accordance with the
statement of Setyaningrum & Wibowo (2016) that the larger the egg diameter, the lower
the fecundity. The size of the egg diameter affected the length of hatching time, where in
the same hatching conditions, the eggs with a larger diameter hatch longer. The same
egg diameter was obtained in T. soro, indicating that the species was the most ready to
spawn and had good egg quality, because T. soro had adapted well to the cultivation
environment compared to the other two species.

Conclusions. Tor tambroides was the species that had the latest gonad maturation with
the highest egg diameter 3.00±0.06 mm and fecundity 1061.19±35.18 eggs kg-1, while
T. douronensis showed significantly lower on fertility and hatching rate with an average
egg diameter of 2.3±0.07 mm and fecundity 4987.80±17.25 eggs kg-1. However, the
survival rate of 15-day-old larvae was not significantly different among species.

Acknowledgements. The authors would like to show appreciation for the Ministry of
Marine Affairs and Fisheries for partly supporting the Thesis Fund. We wish to
acknowledge and thank the Research Institute for Freshwater Aquaculture and Fisheries
Extension (RIFAFE), Bogor, for providing both fish and research facilities. Fish spawning

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was made possible by the excellent support of the RIFAFE staff. No conflict of interest
was to be declared among authors.

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Received: 03 August 2018. Accepted: 02 January 2018. Published online: 28 February 2019.
Authors:
Wahyulia Cahyanti, Research Institute for Freshwater Aquaculture and Fisheries Extension, Jl. Sempur No. 1
Bogor 16129, West Java, Indonesia; Aquaculture Department, Bogor Agricultural University, Dramaga Campus,
Bogor 16680, West Java, Indonesia, e-mail: wahyulia_kkp@yahoo.com
Dinar Tri Soelistyowati, Aquaculture Department, Bogor Agricultural University, Dramaga Campus, Bogor
16680, West Java, Indonesia, e-mail: sdinarts@yahoo.com
Odang Carman, Aquaculture Department, Bogor Agricultural University, Dramaga Campus, Bogor 16680, West
Java, Indonesia, e-mail: odangdoang.od@gmail.com
Anang Hari Kristanto, Research Institute for Freshwater Aquaculture and Fisheries Extension, Jl. Sempur No. 1
Bogor 16129, West Java, Indonesia, e-mail: ananghari25@gmail
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which
permits unrestricted use, distribution and reproduction in any medium, provided the original author and source
are credited.
How to cite this article:
Cahyanti W., Soelistyowati D. T., Carman O., Kristanto A. H., 2019 Artificial spawning and larvae performance
of three Indonesian mahseer species. AACL Bioflux 12(1):280-288.

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